Lipid nanoparticles and preparation methods and use thereof

Abstract

A lipid nanoparticle and a preparation method and use thereof are provided. The lipid nanoparticle (LNP) includes a carrier and an encapsulated nucleic acid, the carrier includes an ionizable lipid, a helper phospholipid, a PEGylated lipid, cholesterol and its derivatives, and a retinoid compound; and the nucleic acid is one or more of mRNA, circRNA, siRNA, microRNA, antisense nucleic acid, and plasmid.

What is claimed is: 
     
         1 . A lipid nanoparticle, wherein the lipid nanoparticle comprises a carrier and an encapsulated nucleic acid, the carrier comprises an ionizable lipid, a helper phospholipid, a PEGylated lipid, cholesterol and derivatives thereof, and a retinoid compound; and the nucleic acid is one or more of mRNA, circRNA, siRNA, microRNA, antisense nucleic acid, and plasmid. 
     
     
         2 . The lipid nanoparticle of  claim 1 , wherein the ionizable lipid accounts for 20 mol %-50 mol % of total lipids in the lipid nanoparticle;
 the ionizable lipid is selected from a group including: 8-[(2-hydroxyethyl) (6-oxo-6-decyloxyhexyl)amino]octanoic acid (heptadecan-9-yl) ester (SM-102), [(4-hydroxybutyl)azetidinyl]bis(hexane-6,1-diyl)bis(2-hexylhexanoate) (ALC-0315), 4(N,N-dimethylamino)butanoic acid (dioleyl) methyl ester (DLin-MC3-DMA), 3,6-bis{4-[bis(2-hydroxydodecyl)amino]butyl}piperazine-2,5-dione (cKK-E12), 9-(4-(dimethylamino)butanoyloxy)heptadecanedioic acid bis((Z)-nonyl-2-en-1-yl) ester (L319), N2,2-dioleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), 8-[(2-hydroxyethyl)(8-nonyloxy-8-oxooctyl)amino]octanoic acid (heptadecan-9-yl) ester (Lipid5), 1,1′-[(2-{4-[2-({2-[bis(2-hydroxydodecyl)amino]ethyl}(2-hydroxydodecyl)amino)ethyl]piperazin-1-yl}ethyl)azabicyclo[bis(dodecane)-2-alcohol) (C12-200), (2,3 dioleoyl-propyl) trimethylammonium chloride (DOTAP), dimethyl dioctadecylammonium bromide (DDAB), and tetrakis(8-methylnonyl) 3,3′,3″,3″-{[(methylazanediyl)bis(propane-3,1diyl)]bis(azatriyl)}tetrapropionate (3060i10); and   the total lipids are a sum of the ionizable lipid, the helper phospholipid, the cholesterol and the derivatives thereof, the PEGylated lipid, and the retinoid compound.   
     
     
         3 . The lipid nanoparticle of  claim 1 , wherein the helper phospholipid accounts for 2 mol %-10 mol % of the total lipids in the lipid nanoparticle; and
 the helper phospholipid is selected from a group including: 1,2-distearoyl-sn-glycero-3-phosphatidylcholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), 2-oleoyl-1-palmitoyl acyl-sn-glycero-3-phosphatidylcholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphatidylethanolamine (POPE), 1,2-distearoyl-sn-glycero-3-phosphatidylethanolamine (DSPE), and 1,2-dipalmitoyl-sn-glycero-3-phosphatidylethanolamine (DPPE).   
     
     
         4 . The lipid nanoparticle of  claim 1 , wherein the cholesterol and the derivatives thereof account for 10 mol %-40 mol % of the total lipids in the lipid nanoparticle; and
 the cholesterol and the derivatives thereof include one or more of β-sitosterol, cholestanol, cholestanone, cholesterol, cholestenone, 7β-hydroxycholesterol, and 7α-hydroxycholesterol.   
     
     
         5 . The lipid nanoparticle of  claim 1 , wherein the PEGylated lipid accounts for  0 . 3  mol %-30 mol % of the total lipids in the lipid nanoparticle; and
 the PEGylated lipid is selected from a group including: 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol (DMG-PEG), 1,2-distearoyl-rac-glycero-3-methoxypolyethylene glycol (DSG-PEG), 1,2-dipalmitoyl-rac-glycero-3-methoxy polyethylene glycol (DPG-PEG), and 1,2-distearoyl-sn-glycero-3-phosphatidylethanolamine-methoxypolyethylene glycol (DSPE-PEG). 
 
     
     
         6 . The lipid nanoparticle of  claim 1 , wherein the retinoid compound accounts for 1 mol %-35 mol % of the total lipids in the lipid nanoparticle; and
 the retinoid compound is one or more of retinoic acid derivatives and synthetic retinoid compounds, the retinoic acid derivatives include one or more of all-trans-retinoic acid (ATRA), 9-cis-retinoic acid (9-cis-RA, 9-CRA), 13-cis-retinoic acid (13-cis-RA, 13-CRA), and the synthetic retinoid compounds include retinoic acid receptor agonist.   
     
     
         7 . The lipid nanoparticle of  claim 1 , wherein a molar ratio of the cholesterol and the derivatives thereof to the retinoid compound is within a range of 1:2-50:1. 
     
     
         8 . The lipid nanoparticle of  claim 1 , wherein an encapsulation efficiency of the nucleic acid is within a range of 30%-99%. 
     
     
         9 . The lipid nanoparticle of  claim 1 , wherein an encapsulation efficiency of the retinoid compound is within a range of 30%-99%. 
     
     
         10 . The lipid nanoparticle of  claim 1 , wherein a molar ratio of nitrogen element of the ionizable lipid to phosphorus element of the nucleic acid is (1-50):1. 
     
     
         11 . The lipid nanoparticle of  claim 1 , wherein a hydrated particle size of the lipid nanoparticle is within a range of 100 nm-200 nm. 
     
     
         12 . A preparation method for the lipid nanoparticle of  claim 1 , comprising: obtaining the lipid nanoparticle based on an aqueous phase containing the nucleic acid and a lipid organic phase by an ethanol dilution method or a microfluidic method. 
     
     
         13 . The preparation method of  claim 12 , wherein the ethanol dilution method comprises: aspirating the aqueous phase containing the nucleic acid into the lipid organic phase, mixing a liquid by rapidly aspirating the liquid with a pipette greater than 50 times, standing after mixing to obtain a product, and obtaining the lipid nanoparticle. 
     
     
         14 . The preparation method of  claim 13 , wherein the ethanol dilution method further comprises dialyzing the product in a 1×PBS buffer solution with a volume of greater than 1000 times of a volume of the product for more than 4 h. 
     
     
         15 . The preparation method of  claim 12 , wherein the microfluidic method comprises: aspirating the lipid organic phase and the aqueous phase containing the nucleic acid with a syringe, respectively, and injecting the lipid organic phase and the aqueous phase into a microfluidic chip at a flow rate of 1:3 for mixing, stopping collecting the liquid after the aqueous phase is empty in the syringe to obtain a collected mixture, and obtaining the lipid nanoparticle. 
     
     
         16 . The preparation method of  claim 15 , wherein the microfluidic method further comprises dialyzing the collected mixture in a 1×PBS buffer solution with a volume of greater than 1000 times of a volume of the mixture for more than 4 h. 
     
     
         17 . A biological product, comprising the lipid nanoparticle of  claim 1 . 
     
     
         18 . The biological product of  claim 17 , wherein the biological product is an injectable biological product; and
 the biological product is a vaccine, preferably an mRNA vaccine.   
     
     
         19 . The biological product of  claim 17 , wherein the biological product is an influenza vaccine, an AIDS (HIV) vaccine, a viral pneumonia (SARS, SARS-COV-2, etc.) vaccine, a tuberculosis vaccine, a norovirus vaccine, a rotavirus (RV) vaccine, a respiratory syncytial virus (RSV) vaccine, an enterovirus (e.g., EV71) vaccine, an intestinal mucosal-related tumor vaccine (e.g., colorectal tumor mRNA vaccine), or a lung cancer vaccine. 
     
     
         20 . The biological product of  claim 17 , wherein the biological product is used for prevention and/or treatment of infectious diseases transmitted via mucosa and mucosa-related tumors; the infectious diseases are selected from influenza, novel coronavirus pneumonia, atypical pneumonia (SARS), tuberculosis, bronchitis and pneumonia caused by RSV infection, AIDS, hand-foot-and-mouth disease caused by enterovirus infection, rotavirus diseases caused by enteroviral infections, and diseases caused by noroviruses; and the mucosa-related tumors are colorectal tumors and lung cancer.